Fig. 1.

Characterization of ASC-CM and -EV. a Representative images of NTA referred to ASC-CM (left) and ASC-EV (right). The table shows the dimensional parameters of the samples expressed as mean ± SD of 6 NTA measurements. b Flow cytometer calibration with standard beads and CFSE⁺ ASC-CM. The FITC⁺ gate encloses the coordinates in SSC-H and FITC-H channels where to expect the events of interest. c–e CD63, CD81, and CD9 staining of representative CFSE⁺ ASC-CM and ASC-EV samples. f Transmission electron microscopy image showing the characteristic morphology of EV. The scale bar indicates 200 nm. g Representative Western blot of ASC-CM and EV lysates deriving from 10⁶ ASC. Cell lysate from 5 × 10⁴ ASC is shown as control. h Laser scanning confocal microscopy of CH treated with ASC^GFP+-EV for 3 days. β-Tubulin was revealed with an Alexa Fluor® 568 conjugated antibody (red), nuclei were stained with DAPI (blue) (magnification × 63). The scale bar indicates 10 μm and the orthogonal views were obtained by Fiji software. i Total protein content per million ASC (μg/10⁶ cells). Data are shown as mean ± SD (n = 4). l Ponceau S staining of ASC-CM and -EV lysates from 10⁶ ASC. Cell lysate from 5 × 10⁴ ASC is also shown